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Recombinant microorganism

a technology of recombinant microorganisms and recombinant proteins, which is applied in the field of recombinant microorganisms, can solve the problems of not necessarily showing high production efficiency of proteins or similar substances in industrial production

Inactive Publication Date: 2014-06-19
KAO CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The patent is about a recombinant microorganism that has been modified by adding a foreign protein or polypeptide gene. This modification is achieved by transferring a mutant strain of microorganism that has certain genes deleted or knocked out, to a new strain. The technical effect of this patent is the creation of a microorganism with a new and useful ability to produce a foreign protein or polypeptide.

Problems solved by technology

However, microorganisms inherently possess diversified genes so that they can cope with environmental changes in the natural world, and thus, they do not necessarily exhibit high production efficiency of proteins or similar substances in industrial production, where only limited production media are employed.

Method used

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  • Recombinant microorganism

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0039]A genome DNA sample, serving as a template, extracted from Bacillus subtilis 168 strain and two primer sets (ccpA-AF and ccpA-A / CmR; and ccpA-B / CmF and ccpA-BR) shown in Table 2 were used to prepare a 0.6 kb fragment (A) flanking the upstream side of the ccpA gene on the genome and a 0.6 kb fragment (B) flanking the downstream side of the ccpA gene. A chloramphenicol-resistant gene of plasmid pC194 (J. Bacteriol. 150 (2), 815 (1982))) was inserted into the XbaI-BamHI cleavages site of plasmid pUC18, to thereby prepare a recombinant plasmid pCBB 31. The recombinant plasmid pCBB and a primer set consisting of CmF and CmR shown in Table 2 were used to prepare a 1 kb fragment (C) containing the chloramphenicol-resistant gene. Subsequently, SOE-PCR was performed by use of the primers ccpA-AF and ccpA-BR shown in Table 2, and by use of the thus-prepared three fragments (A), (B), and (C) in combination as templates, a 2.2 kb DNA fragment in which the fragments (A), (B), and (C) were ...

example 2

[0040]In a manner similar to that described in Example 1, sporulation gene-deleted strains into which a chloramphenicol-resistant gene had been introduced by way of substitution in place of the below-described deleted genes were separated through use of a DNA fragment for effecting deletion prepared from an adequate primer set selected from among various primer sets shown in Table 2; i.e., gene-AF, gene-A / CmR, gene-B / CmF, gene-BR, CmF, and CmR. The gene deleted from the genome was comA, yopO, treR, yvbA, yvaN, yttP, yurK, yozA, licR, sigL, mntR, glcT, ykvE, slr, rocR, yyaA, or rsiX.

example 3

[0041]In a manner similar to that described in Example 2, a DNA fragment for deletion was prepared by use of an adequate primer set selected from among the gene-AF, gene-A / Cm2R, gene-B / Cm2F, gene-BR, Cm2F, and Cm2R, which are shown in Table 2. By use of the thus-prepared DNA fragment, sporulation gene-deleted strains into which a chloramphenicol-resistant gene had been introduced by way of substitution in place of the below-described deleted genes were separated. The gene deleted from the genome was cspB, yvdE, yaaT, yycH, or ylbO.

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Abstract

A recombinant microorganism obtained by transferring, into a host microorganism capable of producing protein or polypeptide with increased productivity, a gene encoding a protein or polypeptide, and a method for producing a protein or polypeptide by use of the recombinant microorganism. The recombinant microorganism is prepared by transferring, to a mutant strain of microorganism from which any of Bacillus subtilis genes comA, yopO, treR, yvbA, cspB, yvaN, yttP, yurK, yozA, licR, sigL, mntR, glcT, yvdE, ykvE, slr, rocR, ccpA, yaaT, yyaA, yycH, yacP, hprK, rsiX, yhdK, and ylbO, or one or more genes functionally equivalent to any of these genes have been deleted or knocked out, a gene encoding a heterologous protein or polypeptide.

Description

TECHNICAL FIELD [0001]The present invention relates to a recombinant microorganism which may be used to produce useful proteins or polypeptides, as well as to such proteins and polypeptides.TECHNICAL BACKGROUND [0002]Microorganisms are widely used for industrially producing a broad range of useful substances, including alcoholic beverages, certain types of foods such as miso and shoyu, amino acids, organic acids, nucleic-acid-related substances, antibiotics, sugars, lipids, and proteins. These substances also find diversified uses, including foods, pharmaceuticals, detergents, products for daily use such as cosmetics, and a variety of chemical raw materials.[0003]In industrial production of useful substances by use of microorganisms, improvement of productivity is one major topic of interest, and one approach therefor is breeding of microorganisms through mutagenesis or other genetic means. Recently, in particular, with advancement of microbial genetics and biotechnology, more effi...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12N15/75C12P21/00C12N15/09C12N1/21C12N9/00C12P21/02
CPCC12N15/75C12P21/00
Inventor TOHATA, MASATOSHISAWADA, KAZUHISAOZAKI, KATSUYAKOBAYASHI, KAZUOOGASAWARA, NAOTAKE
Owner KAO CORP